Applied Food Research 3 (2023) 100309

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Applied Food Research

journal homepage: www.elsevier.com/locate/afres

Making ice cream with natural sweetener stevia: Formulation and

characteristics
Khondoker Shahin Ahmed, G. M. M. Anwarul Hasan∗, Mohammed A. Satter, Kiron Sikdar
Institute of Food Science and Technology (IFST), Bangladesh Council of Scientific and Industrial Research (BCSIR), Dr. Qudrat-E-Khuda Road, Dhaka 1205, Bangladesh

a r t i c l e i n f o a b s t r a c t

Keywords: In the modern era, ice cream is a widely consumed dessert in our society. The main aim of the study is re-
Natural sweetener placement of sugar by natural Sweetener and create good quality of ice cream. Based on chemical, physical, and
Stevia microbiological parameters, ice cream was examined. In our study, we used one formulated ice cream (FIC) and
Sugar
five distinct brand ice creams (A, B, C, D, and E). When sucrose was replaced with stevia, the resulting ice cream
Ice cream
had reduced total solids, fat, ash, freezing point, and hardness (29.43 ± 1.56%; 8.42 ± 0.19%; 0.82 ± 0.07%;
Milk
2.29 ± 0.17 °C; 42.98 ± 3.27 N for FIC and 32.87 ± 1.07 to 35.09 ± 1.18%; 9.90 ± 0.14 to 10.28 ± 0.16%;
3.43 ± 0.13 to 3.67 ± 0.12%; 3.21 ± 0.18 to 3.35 ± 0.10 °C; 43.24 ± 3.57 to 46.21 ± 3.76 N for all brands) but
greater protein, viscocity, and sensory test values (3.74 ± 0.23%; 93.21 ± 1.98 cP; 7.54 for FIC and 2.12 ± 0.08
to 2.40 ± 0.14%; 90.32 ± 1.97 to 92.26 ± 1.29 cP; 6.95 to 7.33 for all brands) when compared to brands that
used sucrose as their sweetener. Among all the brands, FIC sensory acceptability was the highest. We came to
the conclusion that substituting stevia for sucrose may be an option for diabetic patients as well as persons of all
ages.

1. Introduction
Ice cream is currently the most popular frozen food consumed glob-
ally (Alizadeh et al., 2014). A combination of unfrozen components
for example milk, fat, sugar, and flavors are used to make this semi-
solid, cold food product. It now has more significance within the dairy
business and relates to one of the most well-known dairy products
(Akalın et al., 2008). The high calorific value of ice cream is mainly
due to its high level of fat, protein, and carbohydrate. 100 g of decent,
standard ice cream contains 200 kcal of calories. As a result of con-
sumer demand for healthier and more valuable foods, new manufactur-
ing techniques have been developed (Akin et al., 2007; Soukoulis et al.,
2009; Soukoulis & Tzia, 2010). Developing lower-fat, lower-sugar prod-
ucts could boost sales, particularly for frozen sweets (Olson et al., 2003).
Three of the most critical structural elements of ice cream are ice crys-
tals, air cells, and fat corpuscles, all of which are dispersed in a cou-
pled phase from a non-frozen solution (Marshall, 1996). Sweeteners in-
fluence texture, viscosity, and the freezing point making them one of
the ingredients that have a more significant impact on consumer accep-
tance (Alizadeh et al., 2014). Ice cream’s overall composition includes
between 3 and 15% fat, and 9 and 28% sugar (Goff, 2018). Systolic
blood pressure, hypertension, and other conditions are all exacerbated
by sucrose. Additionally, it aggravates diabetes (Deshmukh et al., 2014).
The primary cause of death in most nations and a significant regional
and worldwide public health issue is diabetes (Zimmet et al., 2014). Ac-
cording to the IDF (International Diabetes Federation), there were 465
million (9.3%) cases of diabetes globally in 2019, and by 2045, that
figure might reach 700 million (10.9%) (Atlas, 2019). The prevalence
of adult pre-diabetes was 374 million (7.5%) in 2019 and is projected
to increase to 548 million (8.6%) by 2045, similar to pre-diabetes. On
average, patients with type 2 diabetes mellitus (T2DM) have a 10 year
decline in life expectancy and cardiovascular problems account for 80%
of T2DM patient deaths (Guariguata et al., 2014). Diabetes prevalence
is continuously growing in Bangladesh. In Bangladesh’s International
centre for Diarrhoeal Disease Research, 7.1 million people had diabetes
in 2015; 3.7 million cases went undiagnosed, and the condition was
responsible for about 129,000 fatalities (International Centre, 2019).
According to research that has been published, Bangladesh has a preva-
lence of diabetes that ranges from 2.21%−35% (Akhtar et al., 2020).
Ice cream has been manufactured using a combination of sweeten-
ers. The most popular sweetener in ice cream manufacture is sucrose be-
cause of rheological and financial concerns. It has a high glycemic index,
which is correlated with the metabolic syndrome, type 2 diabetes, obe-
sity, coronary heart disease, hypertension, and dental caries (Aliha et al.,
2013; Alizadeh et al., 2014). Even though these substances produce lit-
tle or no calories, some safety concerns have been highlighted, including

∗
Corresponding author.
E-mail address: pd-cbirmdp@bcsir.gov.bd (G. M. M. Anwarul Hasan).

https://doi.org/10.1016/j.afres.2023.100309
Received 6 April 2023; Received in revised form 15 June 2023; Accepted 16 June 2023
Available online 19 June 2023
2772-5022/© 2023 The Author(s). Published by Elsevier B.V. This is an open access article under the CC BY license (http://creativecommons.org/licenses/by/4.0/)
K.S. Ahmed, G. M. M. Anwarul Hasan, M.A. Satter et al.
Fig. 1. Preparation of Sugar-Free Ice Cream.
their potential for cancer, teratogenicity, and interaction with certain
metabolic or vascular illnesses. As a result, various efforts have been
made to produce foods containing sugar while using natural sweeteners
(Mahan & Escott-Stump, 2013).
In general, sweeteners can be categorized as either natural or syn-
thetic. Depending on the dispersion matrix, natural sweetener stevia
rebaudiana (S. rebaudiana) is 250 to 300 times sweeter than sucrose
(Adari et al., 2016). Those with diabetes mellitus and other seden-
tary lifestyle-related diseases might use it as a healthy alternative to
sugar because of its exceptionally low calorie content (one gram has
no calories) (Curry & Roberts, 2008). The JECFA, WHO, and FDA ac-
knowledge stevia as a safe supplement with rather high maximum limits
(FAO/WHO, 2004; Reddy et al., 2005, 2010).
Since the 1950s, Bangladesh has had an ice cream industry,
which is presently valued at about BDT 65 million (Laskar, 2017;
Humayun, 2018). Due to factors including the industry’s rising growth
rate, the expanding middle class, changing tastes of younger consumers,
the lengthening summer season, and other factors, the market is becom-
ing more and more competitive. (Begum et al., 2020).
Gillespie et al. (2023) showed that, if excess amounts of sugar are
taken by humans then it is responsible for several diseases. Due to the
rise in ice cream manufacturing, the rising number of diabetic patients,
and the harmful effects of sugar on human health, it was determined
to do research on the development of sugar free ice cream that diabetic
patients may consume.
2. Material and methods
2.1. Raw material
The ingredients needed to make ice cream, such as milk powder,
pasteurized milk, sterilized cream, vanilla flavor, and stevia leaf extract,
were purchased from a local market in Dhaka, Bangladesh.
2.2. Packaging material
For the purpose of packaging ice cream, ice cream cups were col-
lected from the Dhaka local market.
2.3. Formulation of ice cream using stevia powder
The ice cream was made in accordance with the flowchart shown
in Fig. 1. Following the recipe, ingredients were brought and weighed.
Table 1 shows how stevia (S. rebaudiana) powder was used to produce
2
Applied Food Research 3 (2023) 100309
Table 1
Formulated Ice cream (g/kg).
Ingredients Sugar free ice cream with Stevia
Milk powder 91.58 (g)
Pasteurized milk 848.90 (g)
Stevia 14.65 (g)
Sterilized cream 36.63 (g)
Vanilla flavor 8.24 (g)
Total 1000 (g)
ice cream in place of sugar. 91.58 g of milk powder and 848.90 g of Pas-
teurized milk were added in the blender (MX-AC400 Mixer Grinder,
220–240 V). We blend those mixtures for 2 min. After that, add 36.63 g
of sterilized cream in a blender. In addition, 8.24 g of vanilla flavor and
14.65 g of stevia leaf extract were added in a blender for 4 min bland.
The mixers were transferred in an Ice cream machine and stay 1 hour.
Finally, we collect Sugar-Free Ice Cream in a cups and stored at freeze.
2.4. Chemical analysis
The several methods were used to investigate the chemical composi-
tion of ice cream. The International Dairy Federation’s 1982 method
was used to assess total solids, the IDF’s 1986 Kjeldhal method was
used to determine protein, ISO 488 was used to measure fat content,
AOAC 2012 to calculate ash content, and ISO 6091 was used to deter-
mine the amount of titrable acidity.
2.5. Physicochemical assessments
Viscocity (NDJ-5S, Drawel, China) was measured by
Atallah et al. (2022), according to Marshall et al. (2003), the freezing
point was determined, a universal testing device was used to evaluate
the hardness of ice cream samples. The speed for hardness test was
adjusted to 60 mm/s while the trigger force was 1 N, 25% deformation
and 2 s of holding time at −20 °C.
2.6. Sugar profile
The profiling of sugar was performed according to
Arslaner et al. (2019) with some modifications. Five-grams of samples
were dissolved in 20 mL of a mixture of water and methanol (75:25;
v/v) and after that, centrifuged at 5000 g for 10 min at 4 °C. Later,
the supernatant was collected and filtered using a 0.45 μm membrane
filter and shifted to 2 mL HPLC vials until further analysis. The samples
were then injected into HPLC system ((UltiMate® 3000 HPLC system;
Dionex, Thermofisher Scientific, USA) consisted of Kromasil 100–5-NH2
column with Heptane as mobile phase and UV detector. The column
oven temperature fixed at 40 °C and finally, 20μL of the extract was
injected. A comparison to sugar standards with retention times was
used to determine the presence of glucose, fructose, galactose, sucrose,
and lactose.
2.7. Microbiological properties
According to Atallah et al. (2022), the total aerobic meshophilic bac-
terial count and the total psychrotrophic bacterial count were assessed.
2.8. Sensory analysis
Five members of a sensory panel, ranging in age from 25 to 45, per-
formed a sensory evaluation of the ice cream samples. The sensory char-
acteristics, such as flavor, body texture, color and appearance, melting
quality, and general acceptance, were evaluated. The 9-point hedonic
scale was used to evaluate the senses (Wichchukit & O’Mahony, 2015).
K.S. Ahmed, G. M. M. Anwarul Hasan, M.A. Satter et al. Applied Food Research 3 (2023) 100309

Table 2
Chemical Analysis%, (g/100 g-1) of sugar free ice cream compared to other available brands in market.

Parameters Ice cream with stevia Brand A Brand B Brand C Brand D Brand E Significance
levels (ANOVA)

Total Solids 29.43 ± 1.56 32.87 ± 1.07 34.39 ± 0.98 35.09 ± 1.18 34.23 ± 1.26 33.98 ± 1.19 0.0013
Protein 3.74 ± 0.23 2.12 ± 0.08 2.19 ± 0.13 2.28 ± 0.09 2.40 ± 0.14 2.35 ± 0.18 <0.0001
Fat 8.42 ± 0.19 9.97 ± 0.18 9.93 ± 0.17 10.28 ± 0.16 10.25 ± 0.12 9.90 ± 0.14 <0.0001
Ash 0.82 ± 0.07 3.43 ± 0.13 3.49 ± 0.21 3.67 ± 0.12 3.54 ± 0.19 3.44 ± 0.22 <0.0001
Titrable acidity 0.19 ± 0.03 0.18 ± 0.08 0.19 ± 0.09 0.18 ± 0.07 0.18 ± 0.07 0.19 ± 0.08 0.9999

The number of sample 3 (n = 3); The results are presented in Mean ± SD.

Table 3
Physical analyses of sugar free ice cream compared to other available brands in market.

Parameters Ice cream with Stevia Brand A Brand B Brand C Brand D Brand E Significance
levels (ANOVA)

Viscocity (cP) 93.21 ± 1.98 91.78 ± 1.19 91.28 ± 1.27 90.32 ± 1.97 91.23 ± 2.01 92.26 ± 1.29 0.4253
Freezing point ( °C) 2.29 ± 0.17 3.32 ± 0.14 3.21 ± 0.18 3.35 ± 0.10 3.30 ± 0.16 3.34 ± 0.17 <0.0001
Hardness (N) 42.98 ± 3.27 46.21 ± 3.48 45.38 ± 4.22 44.97 ± 3.29 46.21 ± 3.76 43.24 ± 3.57 0.7962
Sensory test 7.54 6.95 7.29 7.33 7.12 7.15 –

The number of sample 3 (n = 3); The results are presented in Mean ± SD.

Table 4
Sugar Content of Stevia ice cream compared to other available brands in market% (g 100 g-1).

Parameters Ice cream with Stevia Brand A Brand B Brand C Brand D Brand E Significance
levels (ANOVA)

Glucose 0.23 ± 0.07 0.70 ± 0.11 0.75 ± 0.13 0.72 ± 0.10 0.74 ± 0.12 0.77 ± 0.19 0.0014
Fructose ND 0.22 ± 0.09 0.21 ± 0.09 0.23 ± 0.08 0.22 ± 0.05 0.24 ± 0.08 –
Galactose 0.27 ± 0.08 0.30 ± 0.03 0.29 ± 0.06 0.32 ± 0.04 0.31 ± 0.09 0.29 ± 0.06 0.9431
Sucrose ND 13.98 ± 1.24 12.92 ± 1.09 13.87 ± 1.12 12.98 ± 1.03 13.29 ± 0.98 –
Lactose 3.29 ± 0.47 3.43 ± 0.14 3.68 ± 0.19 3.85 ± 0.12 3.60 ± 0.12 3.73 ± 0.85 0.6093
∗
ND: Not detected; The number of sample 3 (n = 3);.

2.9. Statistical analysis
All experiments were replicated three times. The mean and standard
deviation were used to express the data. The GraphPad Prism 8 was used
to determine the ordinary one-way ANOVA for group comparison and
the Tukey’s multiple comparisons test for pair-wise comparison of FIC
with different brands. A statistically significant level of probability was
defined as ∗ is equal to p < 0.05, ∗∗ is equal to p < 0.01, ∗∗∗ is equal to
p < 0.001, ∗∗∗∗ is equal to p < 0.0001 and ns is equal to nonsignificant.
3. Results and discussions
Using natural sweeteners as an alternative to artificial sugar in ice
cream and examining its chemical, physical, microbiological, and sen-
sory qualities was the main goal of this study. These properties are im-
portant for the quality of ice cream because they have a significant im-
pact on consumers’ consumption of these products. Goff (2018) who
reported ice cream contain between 9 and 28% sugar of the total compo-
nents. Now, People are more conscious of their health state and careful
of their food as a result of the sharp rise in type 2 diabetes in different
age. The manufacture of ice cream is directly impacted by this prob-
lem (Moriano et al., 2017). Due to variations in the quantities of total
solids, protein, fat, and ash in the combinations, the chemical features
of the FIC and different bands of ice cream varied significantly from one
another.
3.1. Chemical analysis
The inclusion of sweeteners has a significant impact on the chemical
properties. Table 2 shows the chemical analysis of the ice cream. The
use of natural sweeteners (stevia) had a significant (p < 0.05) impact on
the changes in total solids, protein, fat, ash, and titrable acidity values
of all samples. Firstly, the total solid in FIC was 29.43 ± 1.56% (w/w),
whereas in different bands which were changed between 32.87 ± 1.07
and 35.09 ± 1.18% (w/w). According to data analysis, fat, ash, freez-
ing point, glucose, fructose, sucrose and lactose of the ice creams were
significantly positive related with the total solids, whereas the protein
and viscocity were negatively linked (Table 6). Total solids of FIC are
significantly (p < 0.01) different in several brands (Fig. 3(A)). The total
amount of solid and dry components in ice cream is known as the total
solids. Alizadeh et al., 2014 found that when stevia was used in place
of sucrose in ice cream, there were substantial alterations in the com-
positional qualities. Deshmukhan et al. (2014) found similar findings,
stating that the presence of stevia reduced the total solid value com-
pared to sugar-added ice cream samples. In addition, protein content
in FIC was 3.74 ± 0.23% but in different bands which were changed
between 2.12 ± 0.08% and 2.40 ± 0.14% (w/w). Protein was signifi-
cantly negative correlated to total solids, fat, ash, freezing point, glu-
cose, fructose and sucrose (Table 6). On the other hand, ptotein of
FIC are significantly (p < 0.0001) different in five brands (Fig. 3(B)).
Alizadeh et al. (2014) was found when stevia used then protein content
also increased. Which is directly correlated our present study. Proteins
assist in emulsification, whipping, and the capacity to hold water, and
they also contribute to the development of the structure of ice cream.
Ice cream manufacturing requires foaming, and milk proteins are known
for their capacity to produce foams. As a result, milk proteins help to
stabilize the air interface in ice cream, which is crucial for the product’s
overall structure and structural stability (Patel et al., 2006). On the other
hand, Fat value was 8.42 ± 0.19% (w/w) in the FIC sample and in dif-
ferent bands which were range from 9.90 ± 0.14 to 10.28 ± 0.16%.
It also seen that, fat was significantly positive and negative connected
to total solids, ash, viscocity, freezing point, glucose, fructose, galac-

3
K.S. Ahmed, G. M. M. Anwarul Hasan, M.A. Satter et al. Applied Food Research 3 (2023) 100309

tose, sucrose and protein Table 6. The fat of FIC are significantly (p <
0.0001) different in all brands (Fig. 3(C)). Deshmukhan et al. (2014) and
Alizadeh et al. (2014) were found that fat content high in control
sample (with sugar) than experiment sample (with stevia) which com-
ply with our study. Fat plays a vital role as a structural agent, helps
to stabilize the air phase, and produces the distinctive sensory qual-
ities that are expected of ice cream (Méndez-Velasco & Goff, 2012;
Rolon et al., 2017). It also affects the release of hydrophobic flavor
molecules (McClements, 2015). Ash content in ICS was 0.82 ± 0.07%
and varied from 3.43 ± 0.13% to 3.67 ± 0.12% (w/w) in all brands.

Fig 2. The representation of developed ice cream features such as chemical analysis, physical analysis, sugar content, and microbial count analysis attributes as
defined by the Principal component 1 (PC1).

4
K.S. Ahmed, G. M. M. Anwarul Hasan, M.A. Satter et al.
Fig 2. Continued
It was significantly positive and negative interacted to total solids, fat,
freezing point, glucose, fructose, sucrose and protein shown in Table 6.
Ash of FIC are significantly (p < 0.0001) different in several brands
(Fig. 3(D)). Lastly, there was no significant difference in titrable acidity
values among all samples (p > 0.05) show in Fig. 3(E) but with aero-
bic meshophilic bacteria and galactose which was significantly positive
and negative interconnected (Table 6). The ice cream with stevia (FIC)
contained the highest amounts of protein but lower value of total solids,
ash, and fat.
3.2. Physical analyses
Viscosity had been regarded as a crucial component of ice cream for-
mulations, and up to a certain extent, it seems necessary for whipping
ability and air retention. A mixture’s viscosity value is influenced by
the presence of fat, protein, stabilizers, bulking agents, and high-quality
components. Table 3 contains a list of all samples’ viscosity levels. The
viscosity level of FIC was 93.21 ± 1.98 cP but in marketed product which
ware ranged from 90.32 ± 1.97 to 92.26 ± 1.29 cP. The FIC showed the
highest amounts of viscosity, whereas the Brand C exhibited the lowest
values. The result reveal that viscosity is significantly positive corre-
lated with fat but it negative associated with total solids and glactose
(Table 6). Fig. 4(A) represent the ANOVA test of viscosity which is not
significantly different (p > 0.05) between FIC and other brands. Accord-
ing to Mayangsari et al. (2019) the stevia addition raised the ice cream
samples’ viscosity levels. The fiber content of the powder stevia leaf was
apparently responsible for the outcome.
Table 3 provides information about the ice cream’s freezing point.
The freezing point of FIC was 2.29 ± 0.17 °C and marketed product
which were almost same (3.21 ± 0.18 to 3.35 ± 0.10 °C). Freezing point
is significantly positive connected with total solids, fat, ash, glucose,
fructose and sucrose (p < 0.05) shown in Table 6. The substitution of
sweeteners and bulking additives for sucrose had an impact on freezing
point levels. On the other hand, a mixture’s freezing point decreases
when there is an increase in protein because it replaces water (Baer &
Keating, 1987). Ptotein of FIC are significantly (p < 0.0001) different of
all brands (Fig. 4(B))
An important aspect to consider is the product’s hardness when it
is at the ideal temperature for scooping or dipping. Many parameters,
such as total solids, the principal melting point, the quantity and kind
5
Applied Food Research 3 (2023) 100309
of stabilizers, etc., have an impact on hardness. As a result, it is op-
timal for all ice creams to have approximately identical overruns and
melting points. When the amount of water-soluble components grew,
melting and freezing values dropped. Because they have an impact on
the melting and freezing points of the mixtures, the amount and type
of ingredients that go into making ice cream must be carefully selected
(Atallah et al., 2022). Table 3 shows the hardness values for sample.
Hardness value of FIC was 42.98 ± 3.27 N but in brands ranged from
43.24 ± 3.57 N to 46.21 ± 3.76 N. The highest hardness value ware de-
tected in the Brand A and D (46.21 ± 3.48 N and 46.21 ± 3.76 N) com-
pared with the other Brands (B, C and E). In contrast, the lowest value
(42.98 ± 3.27 N) was found in ICS. The results reveal no significantly
different (p > 0.05, Fig. 4C) between all ice creams. Due to an increase
in the ratios of total solids, fat, sucrose, and ash in the combinations, all
brands have the maximum value of hardness. The degree of hardness in
ice cream is affected by a number of variables, including overrun, ice
crystal size, fat destabilization, ice phase volume, and the mixture’s con-
sistency properties (Muse & Hartel, 2004). Atallah et al. (2022) found
that the hardness value dropped with the stevia addition compared to
other ice cream samples.
The sensory characteristics of ice cream were evaluated by flavor,
body and texture, color and appearance melting quality and overall
acceptability. The sensory attributes of FIC and five different brands
(A, B, C, D, E) were 7.54, 6.95, 7.29, 7.33, 7.12 and 7.15 respec-
tively. The highest value of sensory attributes was found in FIC (7.54).
Stevia and bulking agents are used to replace sugar in ice cream,
Atallah et al. (2022) found that the sensory scores were more suitable
for ice cream. In order to satisfy consumers, the ice cream market trend
is shifting toward a formulation of ice cream that is free of sugar has
outstanding texture, structure, and sensory qualities.
3.3. Sugar profiles
For FIC and various brands, Table 4 illustrates the sugar levels. Sub-
stitution for sugar with sweeteners for example stevia extract contain
glucose and fructose show significantly positive correlation (Table 6).
In FIC, Glucose content was 0.23 ± 0.07. Whereas in all brands which
ware ranged from 0.70 ± 0.11 to 0.77 ± 0.19%. When compared to
the other samples, the Brand E sample had the highest glucose level
(0.77 ± 0.19%). Fig. 4(D) represent the ANOVA test of glucose which
K.S. Ahmed, G. M. M. Anwarul Hasan, M.A. Satter et al.
Fig 3. ANOVA test for multiple groups of Ice cream (Chemical analysis and Microbiological analyses) and the Tukey’s multiple comparisons test for pair-wise
comparison of FIC with different brands. A statistically significant level of probability was defined as ∗ is equal to p < 0.05, ∗∗ is equal to p < 0.01, ∗∗∗ is equal to p
< 0.001, ∗∗∗∗ is equal to p < 0.0001 and ns is equal to nonsignificant.
is significantly different (p < 0.01) between FIC and five brands In
the FIC, fructose and sucrose levels were not observed but in differ-
ent bands which were range from 0.21 ± 0.09% to 0.24 ± 0.08%
and 12.92 ± 1.09% to 13.98 ± 1.24%, respectively. The highest level
of fructose and sucrose ware obtained in the Brand E and Brand A.
Galactose and Lactose values were not changed significantly (p > 0.05,
Fig. 4E & 4F) in FIC and all Brands (0.27 ± 0.08% to 0.32 ± 0.04% and
3.29 ± 0.47% to 3.85 ± 0.12%). Generally, all Brand (A, B, C, D, and
E) contain the highest values of sucrose, glucose, fructose, galactose,
and lactose whereas, FIC fructose and sucrose were not detected. This
is because sucrose has been completely replaced by sweeteners (stevia).
Sucrose has many disadvantages and it develop several metabolic dis-
eases for example, coronary heart disease, type 2 diabetes mellitus, non-
alcoholic fatty liver disease, Obesity and so on (Gillespie et al., 2023).
For diabetic patients, natural sweetened (Stevia) ice cream may be an al-
ternative. Recent years have seen the rise of a number of healthy trends
6
Applied Food Research 3 (2023) 100309
in dairy products as a result of the rising incidence of diabetes, obesity,
and other health-related problems worldwide. The demand for healthy
products from consumers has sparked innovation and resulted in the cre-
ation of a number of healthy substitutes for ingredients currently used
by the dairy industry (Alizadeh et al., 2014).
3.4. Microbiological analyses
Table 5 shows the total numbers of aerobic mesophilic and psy-
chrotrophic bacteria. Total aerobic mesophilic bacteria counts were sig-
nificantly positive recorded (p > 0.05) with titrable acidity shown in
Table 6. The total number of aerobic mesophilic bacteria is nearly the
same (p > 0.05, Fig. 3F). In FIC was 4.27 ± 0.43 log 10 CFU g-1 but
in all Brands ranged from 4.07 ± 0.39 log 10 CFU g-1 to 4.21 ± 0.48
log 10 CFU g-1. The count of total psychrotrophic bacteria in all sam-
ples ware < 1 log10 CFU g-1. These findings are comparable to those of
K.S. Ahmed, G. M. M. Anwarul Hasan, M.A. Satter et al. Applied Food Research 3 (2023) 100309

Fig 4. ANOVA test for multiple groups of Ice cream (Physical analysis and Sugar content) and the Tukey’s multiple comparisons test for pair-wise comparison of FIC
with different brands. A statistically significant level of probability was defined as ∗ is equal to p < 0.05, ∗∗ is equal to p < 0.01, ∗∗∗ is equal to p < 0.001, ∗∗∗∗ is
equal to p < 0.0001 and ns is equal to nonsignificant.

Table 5
Microbiological analyses (log 10 CFU g-1) of Ice cream with stevia compared to other available ice cream brands in Market.

Bacteria Ice cream with Stevia Brand A Brand B Brand C Brand D Brand E Significance
levels (ANOVA)

Aerobic meshophilic bacteria 4.27 ± 0.43 4.07 ± 0.39 4.19 ± 0.44 4.10 ± 0.54 4.16 ± 0.52 4.21 ± 0.48 0.9952
Psychrotrophic bacteria <1 <1 <1 <1 <1 <1 –

The number of sample 3 (n = 3); The results are presented in Mean ± SD.

Atallah et al. (2022) who investigated the effects of several sweeteners
(sucrose, stevia, sucralose, and sorbitol) on ice cream.
3.5. PCA analysis
Fig. 2 shows Principal component analysis (PCA) was employed in
the present work to figure out the most important characteristics of the
developed and five brand ice cream samples through chemical analysis,
physical analysis, sugar content, microbial count, and sensory analysis.
Based on the multivariate analysis, the first two principal components
described 91.34% of the variations among ice cream samples. Follow-
ing the findings, two PCs presented 91.34% of the variation of the in-
put variables, associated with a significant contraction of the data with
major contributions of 79.19% for the first principle component and
12.15% for the second principle component. In accordance with the con-
tent, PC1 (the main component) was largely attributed to the measure-
ments of total solids, protein, fat, ash, viscocity, freezing point, hardness,
glucose, fructose, galactose, sucrose, lactose, Aerobic meshophilic bac-
teria except titrable acidity, and PC2 (supplementary PC) was associated
to titrable acidity in the ice cream datasets.

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K.S. Ahmed, G. M. M. Anwarul Hasan, M.A. Satter et al. Applied Food Research 3 (2023) 100309

Table 6
Correlation coefficients among Chemical Analysis, Physical analysis, Sugar Content and Microbial Count.

Variables Total Protein Fat Ash Titrable Viscocity Freezing Hardness Glucose Fructose Galactose Sucrose Lactose Aerobic
Solids acidity point meshophilic
bacteria

Total Solids 1
Protein −0.894∗ 1
Fat .956∗∗ −0.940∗∗ 1
Ash .956∗∗ −0.979∗∗ .985∗∗ 1
Titrable acidity −0.392 0.443 −0.594 −0.478 1
Viscocity −0.876∗ 0.743 −0.860∗ −0.803 0.631 1
Freezing point .927∗∗ −0.972∗∗ .974∗∗ .992∗∗ −0.495 −0.752 1
Hardness 0.525 −0.675 0.690 0.643 −0.744 −0.661 0.605 1
Glucose .943∗∗ −0.968∗∗ .955∗∗ .988∗∗ −0.360 −0.722 .983∗∗ 0.572 1
Fructose .934∗∗ −0.971∗∗ .966∗∗ .991∗∗ −0.437 −0.735 .998∗∗ 0.569 .991∗∗ 1
Galactose 0.806 −0.707 .876∗ 0.789 −0.834∗ −0.916∗ 0.781 0.664 0.699 0.749 1
Sucrose .925∗∗ −0.989∗∗ .971∗∗ .995∗∗ −0.486 −0.764 .996∗∗ 0.641 .983∗∗ .993∗∗ 0.767 1
Lactose .904∗ −0.669 0.763 0.766 −0.160 −0.775 0.740 0.145 0.763 0.761 0.675 0.723 1
Aerobic meshophilic −0.572 0.744 −0.735 −0.703 .846∗ 0.710 −0.719 −0.766 −0.602 −0.674 −0.804 −0.737 −0.329 1
bacteria

Conclusion
The qualities of the ice cream could be affected by how it is made
from different components. As a result, it should be understood that the
formulation and ingredient selection are crucial to the success of ice
cream production. The impact of sweeteners (stevia) on the chemical,
physical, microbiological, and sensory aspects of ice cream was inves-
tigated in this study. On the other hand, compared FIC with other five
different brands (A, B, C, D, and E). Using stevia as a natural sweet-
ener has a favorable effect on the creation of ice cream. The results of
the current study suggest that selling ice cream to people with diabetes
could be successful if natural sweeteners (stevia) are used. Hence, the
ice cream market is shifting toward a sizable market for sugar free ice
cream as well as formulations with outstanding texture, structure, and
sensory qualities to obtain consumer pleasure. The risk of diseases may
be decreased if natural sweeteners are used in place of sucrose when
making ice cream.
Ethical statement
The authors declare that the material is the authors’ own original
work, which has not been previously published elsewhere. The work is
not currently being considered for publication elsewhere. The partici-
pants provided their oral consent to participate in this study.
Declaration of Competing Interest
The authors declare no conflicts of interest.
Data availability
The data that has been used is confidential.
Acknowledgments
We greatly acknowledge the authority of the Bangladesh Council of
Scientific and Industrial Research (BCSIR) for providing the necessary
lab space and instrument necessary for this research.
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